Fragment-Based Methodology for the Identification of Phosphatase Inhibitors

描述（由申请方提供）：本提案旨在开发基于片段的方法学，以鉴定磷酸酶抑制剂。拟开发的方法包括三个步骤：（1）筛选具有不同的低分子量O-芳基的O-芳基磷酸酯的文库，以使用简单的基于荧光定量的测定来鉴定磷酸酶底物，（2）通过快速类似物合成和评估来优化鉴定的O-芳基磷酸酯底物，和（3）通过用已知的磷酸盐电子等排体直接置换磷酸盐，将优化的底物转化为抑制剂。为了开发该方法，蛋白酪氨酸1B（PTP1B），一种与糖尿病有关的磷酸酶，将被靶向。磷酸酶已被证明可以增强几种疾病的疾病状态，包括骨质疏松症、免疫疾病、癌症、阿尔茨海默病、糖尿病和肥胖症。特别地，PTP1B是治疗2型糖尿病的充分验证的靶标。PTP1B的抑制已被证明有助于治疗小鼠糖尿病而无副作用。虽然PTP1B正在被追求作为一个初始目标，该方法将适用于大量的磷酸酶参与各种疾病。使用常用的组合方法发现PTP1B抑制剂并非没有困难。特别麻烦的是在针对PTP1B的筛选中获得的大量假阳性。所提出的方法将直接解决与磷酸酶抑制剂设计的替代方法发现的陷阱。所提出的方法将能够生成针对磷酸酶家族的抑制剂。磷酸酶与许多疾病有关，包括骨质疏松症、免疫疾病、癌症、阿尔茨海默病、糖尿病和肥胖症。最初的努力将集中在开发与2型糖尿病有关的磷酸酶抑制剂上。由这种方法产生的磷酸酶抑制剂可以为开发针对许多严重疾病的药物提供起点。

DESCRIPTION (provided by applicant): This proposal aims to develop fragment-based methodology toward the identification of phosphatase inhibitors. The method to be developed consists of three steps: (1) a library of O-aryl phosphates with diverse, low molecular weight O-aryl groups is screened to identify phosphatase substrates using a simple spectrophotometric-based assay, (2) the identified O-aryl phosphate substrates are optimized by rapid analogue synthesis and evaluation, and (3) the optimized substrates are converted to inhibitors by direct replacement of the phosphate with known phosphate isosteres. To develop the methodology, protein tyrosine 1B (PTP1B), a phosphatase implicated in diabetes, will be targeted. Phosphatases have been shown to enhance disease states in several maladies including osteoporosis, immune diseases, cancer, Alzheimer's disease, diabetes, and obesity. In particular, PTP1B is a well- validated target in the treatment of type 2 diabetes. Inhibition of PTP1B has been shown to aid in the treatment of diabetes in mice without side effects. While PTP1B is being pursued as an initial target, the methodology will be applicable to a large number of phosphatases involved in a variety of diseases. Finding inhibitors of PTP1B using commonly adopted combinatorial methods has not been without difficulty. Particularly troublesome is the high number of false positives obtained in screens against PTP1B. The methodology proposed will directly address the pitfalls found with alternative methodologies toward phosphatase inhibitor design. The proposed methodology will enable the generation of inhibitors toward families of phosphatase enzymes. Phosphatases have been implicated in a number of diseases including osteoporosis, immune diseases, cancer, Alzheimer's disease, diabetes, and obesity. Initial efforts will be focused on development of inhibitors toward a phosphatase involved in type 2 diabetes. Phosphatase inhibitors generated from this methodology could provide starting points for the development of Pharmaceuticals against a number of serious maladies.